KLF4调控食管鳞癌细胞系TE1奥沙利铂耐药性的机制研究

The mechanism of KLF4 regulating oxaliplatin resistance in esophageal squamous cell carcinoma cell line TE1

目的探讨KLF4对食管鳞癌(ESCC)细胞系TE1奥沙利铂(OXA)耐药性的作用及可能机制。方法OXA浓度梯度筛选法诱导建立TE1/OXA耐药细胞,MTT方法鉴定细胞耐药结果;qRT-PCR检测TE1/OXA耐药细胞KLF4 mRNA表达。构建KLF4过表达耐药细胞TE1/OXA+KLF4及其阴性对照细胞TE1/OXA+cmv,流式细胞术检测转染后细胞凋亡率与阿霉素(ADR)干预后细胞ADR泵出率变化;划痕实验检测各组细胞迁移率;Western blot和qRT-PCR分别检测各组细胞内ABCG2、Nanog、MDR1蛋白与mRNA的表达变化。结果TE1/OXA细胞构建成功,TE1/OXA细胞KLF4表达水平显著低于TE1细胞;过表达KLF4能够显著促进耐药细胞凋亡、抑制体外细胞迁移率及ADR泵出率;TE1/OXA+KLF4细胞中ABCG2、Nanog、MDR1蛋白及mRNA表达水平均较TE1/OXA+cmv组细胞显著减少。结论KLF4可能通过抑制细胞内ABCG2、Nanog、MDR1表达逆转食管鳞癌细胞系TE1奥沙利铂耐药性。

Objective To investigate the effect of KLF4 on the resistance of esophageal squamous cell carcinoma(ESCC)cell line TE1 to oxaliplatin(OXA)effects and possible mechanisms.Methods TE1/OXA-resistant cells were induced and established by OXA concentration gradient screening method,and the results of cell resistance were identified by MTT method.The expression of KLF4 mRNA in TE1/OXA-resistant cells was detected by qRT-PCR.KLF4-overexpressing drug-resistant cells TE1/OXA+KLF4 and negative control cells TE1/OXA+cmv were constructed,and flow cytometry was used to detect the apoptosis rate after transfection and the change of ADR pumping rate in cells after doxorubicin(ADR)intervention.Scratch test was used to detect the cell migration rate in each group.Western blot and qRT-PCR was used to detect the levels of ABCC2,Nanog,MDR1 protein and mRNA in each group respectively.Results TE1/OXA cells were successfully constructed,and the level of KLF4 in TE1/OXA cells was significantly lower than that in TEI cells.Overexpression of KLF4 could significantly promote the apoptosis of drug-resistant cells,inhibit cell migration rate and ADR pumping rate in vitro.Compared with TE1/OXA+cmv group,the levels of ABCG2,Nanog,MDR1 protein and mRNA were significantly decreased in TE1/OXA+KLF4 cells.Conclusion KLF4 may reverse the oxaliplatin resistance of esophageal squamous cell carcinoma cell line TE1 by inhibiting the expressions of ABCG2,Nanog and MDR1.